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  • Title: Development and validation of spectrophotometric and high-performance column liquid chromatographic methods for the simultaneous determination of dienogest and estradiol valerate in pharmaceutical preparations.
    Author: Cağlayan MG, Palabiyik IM, Onur F.
    Journal: J AOAC Int; 2010; 93(3):862-8. PubMed ID: 20629388.
    Abstract:
    Simultaneous determination of dienogest (DIE) and estradiol valerate (EST) in sugar-coated tablets was performed by using HPLC and spectrophotometry. In HPLC, the separation was achieved on an ACE C8 column using the mobile phase acetonitrile-NH4NO3 (0.03 M, pH 5.4; 70 + 30, v/v) at a flow rate of 2 mL/min. The detection wavelength was 280 nm, and cyproterone acetate was selected as an internal standard. The linearity range was 3.0-45.0 microg/mL for DIE and 18.0-100.0 microg/mL for EST. As spectrophotometric methods, two chemometric methods, principal component regression and partial least-squares, were developed. In the chemometric techniques, the concentration data matrix was prepared by using mixtures containing these drugs in methanol-water (3 + 1, v/v). The absorbance data matrix corresponding to the concentration data matrix in these methods was obtained by the measurement of absorbances in their zero-order spectra; then, the calibration was obtained by using the data matrix for the prediction of unknown concentrations of DIE and EST in their binary mixture. Working ranges were found as 2.0-24.0 microg/mL for DIE and 20.0-270.0 microg/mL EST in the methods. These three developed methods were validated and successfully applied to a pharmaceutical preparation, a sugar-coated tablet, and the results were compared with each other.
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